IGEM:British Columbia/Protocols/Amplifying BioBrick Parts: Difference between revisions
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(New page: ==Amplifying Biobrick Parts== ===Materials=== * 2009 Spring Distribution plates * Competent cells * sdH2O * LB agar plates (with appropriate antibiotics) * LB broth (with appropriate anti...) |
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* Resuspend in a PCR tube with 15uL of sdH2O (by pipetting in and out of the water). | * Resuspend in a PCR tube with 15uL of sdH2O (by pipetting in and out of the water). | ||
* [[IGEM:British Columbia/Protocols/Transformation Competent Cells| Transform]] competent cells with 1uL of the resuspended DNA, plate bacteria with the appropriate antibiotic, and grow overnight. | * [[IGEM:British Columbia/Protocols/Transformation Competent Cells| Transform]] competent cells with 1uL of the resuspended DNA, plate bacteria with the appropriate antibiotic, and grow overnight. | ||
* Pick a single colony and [[IGEM:British Columbia/Protocols/Day/Overnight Culture |inoculate]] ~3.5-6mL of medium (with the correct antibiotic) and grow overnight (16-18 hours). | |||
* Make a [[IGEM:British Columbia/Protocols/Glycerol Stocks| glycerol stock]] using 0.75mL of culture. | |||
* [[iGEM:British Columbia/Protocols/Miniprep| Miniprep]] the DNA using ~2.5-5mL of culture | |||
* Label well and store at -4°C. |
Latest revision as of 15:23, 7 August 2009
Amplifying Biobrick Parts
Materials
- 2009 Spring Distribution plates
- Competent cells
- sdH2O
- LB agar plates (with appropriate antibiotics)
- LB broth (with appropriate antibiotics)
Equipment
- miniprep kits (1 prep/part)
Method
- With a pipette tip, punch a hole through the foil cover into the well corresponding to the part you want. Make sure that the plate is properly oriented
- Resuspend in a PCR tube with 15uL of sdH2O (by pipetting in and out of the water).
- Transform competent cells with 1uL of the resuspended DNA, plate bacteria with the appropriate antibiotic, and grow overnight.
- Pick a single colony and inoculate ~3.5-6mL of medium (with the correct antibiotic) and grow overnight (16-18 hours).
- Make a glycerol stock using 0.75mL of culture.
- Miniprep the DNA using ~2.5-5mL of culture
- Label well and store at -4°C.